This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The emerging fields of metagenomics and proteomics are significantly transforming our understanding of the roles microorganisms play in all ecosystems, from the human biome to toxin laden environments. Microbiology is no longer restricted to analysis of pure cultures of one species, isolated from natural habitats and interactions with other microflora. The long range goal of this investigation is to assess the microbial community of a complex, environmentally-stressed site within Lamoille County, Vermont. The Vermont Asbestos Group Mine site, located in Lowell and Eden is impacting both the environmental and human health of the region (Levy, 2008;Vermont Dept of Health, 2008). Water and stream bed quality are contaminated with heavy metals, elevated pH levels, and asbestos-laden mine tailings. an A health advisory indicates increased potential risk to humans of asbestos-related illnesses. The objectives of the project are to (a) construct a metagenomic library of community DNA from an impacted aquatic environment at the Mine site, (b) create a cDNA library of expressed genes of the microorganisms inhabiting the site, and (c) isolate total protein from the microbial community for future proteomic analysis. This integrative approach will enable us to address the dynamic roles microorganisms play in complex, human-impacted environments.